Regulating Gene Expression With Light-Activated Oligonucleotides

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dc.contributor.advisorIvan J. Dmochowski
dc.contributor.authorGriepenburg, Julianne C
dc.date2023-05-17T12:51:38.000
dc.date.accessioned2023-05-22T16:29:51Z
dc.date.available2015-11-24T00:00:00Z
dc.date.copyright2015-11-16T00:00:00-08:00
dc.date.issued2014-01-01
dc.date.submitted2015-11-16T13:06:36-08:00
dc.description.abstractThe work in this thesis identifies new photochemical approaches to gain high spatiotemporal control over molecular structure and function, for broad applications in materials and biological science. "Caged" compounds provide a method for temporarily blocking function until acted upon by an external trigger, typically near-UV light. To enable multiplexing studies, three new biomolecular caging strategies were developed that can be activated with various wavelengths of near-UV or visible light. The first method, an oligonucleotide hairpin structure incorporating one or two nitrobenzyl photolinkers, was applied to a miRNA antagomir and used to "turn off" let-7 miRNA in zebrafish embryos with 365 nm light. To achieve bidirectional control over miRNA, a circular construct was designed for the ability to "turn on" the release of exogenous miRNA into zebrafish embryos with 365 nm light. A second oligonucleotide caging method, using a ruthenium-based photolinker (RuBEP), was designed to extend photoactivation to the visible spectrum, with additional potential for two-photon activation. RuBEP was used to cage antisense morpholinos through circularization via a Cu(I)-mediated [3+2] Huisgen cycloaddition reaction. RuBEP-caged morpholinos were photoactivated to "turn on" antisense activity and successfully knocked down zebrafish chd and ntl genes with 450 nm light, with limited background activity prior to irradiation. A third method of caging was based on encapsulation within photoresponsive nano-polymersomes. Self-assembly of nano-polymersomes was optimized to generate visible-light-responsive vesicles that incorporate a porphyrin dimer in the hydrophobic membrane. These nanovesicles were shown to encapsulate a variety of cargo, including 25mer oligonucleotides, a small molecule fluorescent dye, and two biologically relevant metal ions, Zn2+ and Ca2+. The photoresponsiveness of the system was modulated with light wavelength, irradiation time, and the presence of dextran in the aqueous core.
dc.description.degreeDoctor of Philosophy (PhD)
dc.format.extent229 p.
dc.format.mimetypeapplication/pdf
dc.identifier.urihttps://repository.upenn.edu/handle/20.500.14332/28093
dc.languageen
dc.legacy.articleid3109
dc.legacy.fulltexturlhttps://repository.upenn.edu/cgi/viewcontent.cgi?article=3109&context=edissertations&unstamped=1
dc.provenanceReceived from ProQuest
dc.rightsJulianne C. Griepenburg
dc.source.issue1297
dc.source.journalPublicly Accessible Penn Dissertations
dc.source.statuspublished
dc.subject.otherLight-activated
dc.subject.otherOligonucleotides
dc.subject.otherPhoto-activated
dc.subject.otherPolymersomes
dc.subject.otherRuthenium
dc.subject.otherZebrafish
dc.subject.otherBiochemistry
dc.subject.otherChemistry
dc.titleRegulating Gene Expression With Light-Activated Oligonucleotides
dc.typeDissertation/Thesis
digcom.contributor.authorisAuthorOfPublication|email:jewel1231@gmail.com|institution:University of Pennsylvania|Griepenburg, Julianne C
digcom.date.embargo2015-11-24T00:00:00-08:00
digcom.identifieredissertations/1297
digcom.identifier.contextkey7851098
digcom.identifier.submissionpathedissertations/1297
digcom.typedissertation
dspace.entity.typePublication
relation.isAuthorOfPublicatione8bca80b-0f64-420d-b5e7-e47a952b33b6
relation.isAuthorOfPublication.latestForDiscoverye8bca80b-0f64-420d-b5e7-e47a952b33b6
upenn.graduate.groupChemistry
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